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The intermediate filament protein peripherin is the specific interaction partner of mouse BPAG1-n (dystonin) in neurons.

Leung CL, Sun D, Liem RK - J. Cell Biol. (1999)

Bottom Line: Kouklis, D.W.Fuchs. 1996.Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

View Article: PubMed Central - PubMed

Affiliation: Departments of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York 10032, USA.

ABSTRACT
The dystonia musculorum (dt) mouse suffers from severe degeneration of primary sensory neurons. The mutated gene product is named dystonin and is identical to the neuronal isoform of bullous pemphigoid antigen 1 (BPAG1-n). BPAG1-n contains an actin-binding domain at its NH2 terminus and a putative intermediate filament-binding domain at its COOH terminus. Because the degenerating sensory neurons of dt mice display abnormal accumulations of intermediate filaments in the axons, BPAG1-n has been postulated to organize the neuronal cytoskeleton by interacting with both the neurofilament triplet proteins (NFTPs) and microfilaments. In this paper we show by a variety of methods that the COOH-terminal tail domain of mouse BPAG1 interacts specifically with peripherin, but in contrast to a previous study (Yang, Y., J. Dowling, Q.C. Yu, P. Kouklis, D.W. Cleveland, and E. Fuchs. 1996. Cell. 86:655-665), mouse BPAG1 fails to associate with full-length NFTPs. The tail domains interfered with the association of the NFTPs with BPAG1. In dt mice, peripherin is present in axonal swellings of degenerating sensory neurons in the dorsal root ganglia and is downregulated even in other neural regions, which have no obvious signs of pathology. Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

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Schematic representation of  the mouse BPAG1 COOH-terminal  proteins. The top line corresponds to  the partial mouse BPAG1 cDNA with  the restriction sites used in this study.  The arrows underneath this line denote  the primers used for RT-PCR. Diagrammatic representations of FLAG-tagged BPAG1 COOH-terminal proteins used for transient transfection  assays and GBD-fused mBPAG-C1  and mBPAG-r220 used for two-hybrid  studies are shown. The numbers of the  amino acids are assigned according to  the human BPAG1-e sequence. Compared with that of the human orthologue, the tail domain of mouse  BPAG1 has an extra glutamine before  the stop codon. B and C stand for the B  and C subdomains of BPAG1, respectively. The shaded box symbolizes the  COOH terminus of the mouse BPAG1  rod domain.
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Figure 1: Schematic representation of the mouse BPAG1 COOH-terminal proteins. The top line corresponds to the partial mouse BPAG1 cDNA with the restriction sites used in this study. The arrows underneath this line denote the primers used for RT-PCR. Diagrammatic representations of FLAG-tagged BPAG1 COOH-terminal proteins used for transient transfection assays and GBD-fused mBPAG-C1 and mBPAG-r220 used for two-hybrid studies are shown. The numbers of the amino acids are assigned according to the human BPAG1-e sequence. Compared with that of the human orthologue, the tail domain of mouse BPAG1 has an extra glutamine before the stop codon. B and C stand for the B and C subdomains of BPAG1, respectively. The shaded box symbolizes the COOH terminus of the mouse BPAG1 rod domain.

Mentions: To study the interactions of BPAG1 with nIFs, we isolated three consecutive overlapping pieces of the BPAG1 cDNA by RT-PCR on mouse brain mRNA. The three amplified DNA fragments were ligated together to yield a ∼3.0-kb partial mouse BPAG1 cDNA. Sequencing of this cDNA showed that it encoded 988 amino acids, including the entire tail domain (768 amino acids) and the COOH terminus of the rod domain (220 amino acids). Because the complete mouse BPAG1 cDNA sequence had not been fully characterized (Amagai et al., 1990), the nomenclature of the mouse BPAG1 COOH-terminal proteins was designated according to the sequence of the human BPAG1 epidermal isoform (Fig. 1). We compared our sequence with several EST clones using BLAST. The sequences of a series of I.M.A.G.E. clones (clones 1262090, 931676, 1227492, 1242939, 975299, 317229, and 975617) are identical to the sequence of mBPAG-C2 and cover 75% of the sequence. Furthermore, the mBPAG-C1 sequence is identical to the deduced protein sequence of BPM1, a partial mouse BPAG1 cDNA (accession number 321215; Amagai et al., 1990). The accession number for the mouse BPAG-C1 sequence is AF115383.


The intermediate filament protein peripherin is the specific interaction partner of mouse BPAG1-n (dystonin) in neurons.

Leung CL, Sun D, Liem RK - J. Cell Biol. (1999)

Schematic representation of  the mouse BPAG1 COOH-terminal  proteins. The top line corresponds to  the partial mouse BPAG1 cDNA with  the restriction sites used in this study.  The arrows underneath this line denote  the primers used for RT-PCR. Diagrammatic representations of FLAG-tagged BPAG1 COOH-terminal proteins used for transient transfection  assays and GBD-fused mBPAG-C1  and mBPAG-r220 used for two-hybrid  studies are shown. The numbers of the  amino acids are assigned according to  the human BPAG1-e sequence. Compared with that of the human orthologue, the tail domain of mouse  BPAG1 has an extra glutamine before  the stop codon. B and C stand for the B  and C subdomains of BPAG1, respectively. The shaded box symbolizes the  COOH terminus of the mouse BPAG1  rod domain.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2132913&req=5

Figure 1: Schematic representation of the mouse BPAG1 COOH-terminal proteins. The top line corresponds to the partial mouse BPAG1 cDNA with the restriction sites used in this study. The arrows underneath this line denote the primers used for RT-PCR. Diagrammatic representations of FLAG-tagged BPAG1 COOH-terminal proteins used for transient transfection assays and GBD-fused mBPAG-C1 and mBPAG-r220 used for two-hybrid studies are shown. The numbers of the amino acids are assigned according to the human BPAG1-e sequence. Compared with that of the human orthologue, the tail domain of mouse BPAG1 has an extra glutamine before the stop codon. B and C stand for the B and C subdomains of BPAG1, respectively. The shaded box symbolizes the COOH terminus of the mouse BPAG1 rod domain.
Mentions: To study the interactions of BPAG1 with nIFs, we isolated three consecutive overlapping pieces of the BPAG1 cDNA by RT-PCR on mouse brain mRNA. The three amplified DNA fragments were ligated together to yield a ∼3.0-kb partial mouse BPAG1 cDNA. Sequencing of this cDNA showed that it encoded 988 amino acids, including the entire tail domain (768 amino acids) and the COOH terminus of the rod domain (220 amino acids). Because the complete mouse BPAG1 cDNA sequence had not been fully characterized (Amagai et al., 1990), the nomenclature of the mouse BPAG1 COOH-terminal proteins was designated according to the sequence of the human BPAG1 epidermal isoform (Fig. 1). We compared our sequence with several EST clones using BLAST. The sequences of a series of I.M.A.G.E. clones (clones 1262090, 931676, 1227492, 1242939, 975299, 317229, and 975617) are identical to the sequence of mBPAG-C2 and cover 75% of the sequence. Furthermore, the mBPAG-C1 sequence is identical to the deduced protein sequence of BPM1, a partial mouse BPAG1 cDNA (accession number 321215; Amagai et al., 1990). The accession number for the mouse BPAG-C1 sequence is AF115383.

Bottom Line: Kouklis, D.W.Fuchs. 1996.Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

View Article: PubMed Central - PubMed

Affiliation: Departments of Biochemistry and Molecular Biophysics, Columbia University College of Physicians and Surgeons, New York 10032, USA.

ABSTRACT
The dystonia musculorum (dt) mouse suffers from severe degeneration of primary sensory neurons. The mutated gene product is named dystonin and is identical to the neuronal isoform of bullous pemphigoid antigen 1 (BPAG1-n). BPAG1-n contains an actin-binding domain at its NH2 terminus and a putative intermediate filament-binding domain at its COOH terminus. Because the degenerating sensory neurons of dt mice display abnormal accumulations of intermediate filaments in the axons, BPAG1-n has been postulated to organize the neuronal cytoskeleton by interacting with both the neurofilament triplet proteins (NFTPs) and microfilaments. In this paper we show by a variety of methods that the COOH-terminal tail domain of mouse BPAG1 interacts specifically with peripherin, but in contrast to a previous study (Yang, Y., J. Dowling, Q.C. Yu, P. Kouklis, D.W. Cleveland, and E. Fuchs. 1996. Cell. 86:655-665), mouse BPAG1 fails to associate with full-length NFTPs. The tail domains interfered with the association of the NFTPs with BPAG1. In dt mice, peripherin is present in axonal swellings of degenerating sensory neurons in the dorsal root ganglia and is downregulated even in other neural regions, which have no obvious signs of pathology. Since peripherin and BPAG1-n also display similar expression patterns in the nervous system, we suggest that peripherin is the specific interaction partner of BPAG1-n in vivo.

Show MeSH
Related in: MedlinePlus